GSM8047334: C. difficile, Commercial Porcine Gastric Mucus (PGM, Condition 2), Rep B; Clostridioides difficile R20291; RNA-Seq
C. difficile RNA extracted by bead beating in 1 mL TriZol, followed by addition of 200 µl chloroform. Samples centrifuged, then aqueous layers containing RNA were mixed 1:1 with 70% cold ethanol. This mixture was then applied to columns in an RNeasy kit (Qiagen) and purified using manufacturer instructions, including on-column DNase treatment with RNase-Free DNase Set (Qiagen) prior to elution. Libraries prepared using GENEWIZ (now Azenta Life Sciences) commercial services. Library preparation included RNA quality assessment, rRNA depletion, and adapter ligation (all completed by vendor).